High-resolution transport-of-intensity quantitative phase microscopy with annular illumination

For quantitative phase imaging (QPI) based on transport-of-intensity equation (TIE), partially coherent illumination provides speckle-free imaging, compatibility with brightfield microscopy, and transverse resolution beyond coherent diffraction limit. Unfortunately, in a conventional microscope with circular illumination aperture, partial coherence tends to diminish the phase contrast, exacerbating the inherent noise-to-resolution tradeoff in TIE imaging, resulting in strong low-frequency artifacts and compromised imaging resolution. Here, we demonstrate how these issues can be effectively addressed by replacing the conventional circular illumination aperture with an annular one. The matched annular illumination not only strongly boosts the phase contrast for low spatial frequencies, but significantly improves the practical imaging resolution to near the incoherent diffraction limit. By incorporating high-numerical aperture (NA) illumination as well as high-NA objective, it is shown, for the first time, that TIE phase imaging can achieve a transverse resolution up to 208 nm, corresponding to an effective NA of 2.66. Time-lapse imaging of in vitro Hela cells revealing cellular morphology and subcellular dynamics during cells mitosis and apoptosis is exemplified. Given its capability for high-resolution QPI as well as the compatibility with widely available brightfield microscopy hardware, the proposed approach is expected to be adopted by the wider biology and medicine community.Comment: This manuscript was originally submitted on 20 Feb. 201

Computational Depth-resolved Imaging and Metrology

In this thesis, the main research challenge boils down to extracting 3D spatial information of an object from 2D measurements using light. Our goal is to achieve depth-resolved tomographic imaging of transparent or semi-transparent 3D objects, and to perform topography characterization of rough surfaces. The essential tool we used is computational imaging, where depending on the experimental scheme, often indirect measurements are taken, and tailored algorithms are employed to perform image reconstructions. The computational imaging approach enables us to relax the hardware requirement of an imaging system, which is essential when using light in the EUV and x-ray regimes, where high-quality optics are not readily available. In this thesis, visible and infrared light sources are used, where computational imaging also offers several advantages. First of all, it often leads to a simple, flexible imaging system with low cost. In the case of a lensless configuration, where no lenses are involved in the final image-forming stage between the object and the detector, aberration-free image reconstructions can be obtained. More importantly, computational imaging provides quantitative reconstructions of scalar electric fields, enabling phase imaging, numerical refocus, as well as 3D imaging

Interferometric Synthetic Aperture Microscopy (ISAM) Reconstruction and Characterization in a High Numerical Aperture System

Optical coherence microscopy (OCM) is an imaging modality that is capable of visualizing structural features of biological samples at high resolution based on their scattering properties. Interferometric synthetic aperture microscopy (ISAM) is a newer technique that can overcome the typical dependence between lateral resolution and depth-of-focus of an optical coherence tomography (OCT) imaging system by offering spatially invariant resolution within the whole 3D data set, including regions that are outside of the focal region. Both OCM and ISAM have many potential research and clinical applications. By combining OCM and ISAM, it is possible to visualize an entire 3D volumetric data set with the high resolution normally available only at the focus. Therefore, this combination will yield more detailed information from the observed sample than OCM alone. This combination will also improve the feasibility of the ISAM technique for wider research and clinical applications. This thesis presents the experimental validation and characterization of ISAM applied to high numerical aperture OCM optical imaging. The validation includes the image reconstruction of a tissue phantom containing nano-particles both for OCT and ISAM, and system characterization includes quantitative assessment of the confocal parameter, point spread function, and phase stability measurements. Several potential applications also are examined as a part of this thesis

Single-shot compressed ultrafast photography: a review

Compressed ultrafast photography (CUP) is a burgeoning single-shot computational imaging technique that provides an imaging speed as high as 10 trillion frames per second and a sequence depth of up to a few hundred frames. This technique synergizes compressed sensing and the streak camera technique to capture nonrepeatable ultrafast transient events with a single shot. With recent unprecedented technical developments and extensions of this methodology, it has been widely used in ultrafast optical imaging and metrology, ultrafast electron diffraction and microscopy, and information security protection. We review the basic principles of CUP, its recent advances in data acquisition and image reconstruction, its fusions with other modalities, and its unique applications in multiple research fields

Design and evaluation of a full-sensitivity tilt scanning interferometry system for displacement field tomography and profilometry

This thesis reports the investigation and further development of a tilt scanning interferometry system for surface profilometry and sub-surface tomographic applications. A new 3D full sensitivity interferometry system extends the work carried out on a previous prototype that was capable of measuring displacement along one lateral plus the axial component. Depth-resolved imaging is achieved by the acquisition of a sequence of 2D interferograms whilst the illumination beam undergoes a constant rate of tilt and full sensitivity displacement is achieved by performing scans from multiple illumination directions. The comparison of phase volumes from two successive series of scans enables 3D displacement fields to be determined. The working principle that describes the technique is presented, covering the reconstruction of a depth-resolved sample from the detected intensity distribution. The system performance is studied, including measurement repeatability and factors that affect the depth resolution and depth range. Depth resolution is fundamentally limited by the range of the illumination tilting angle and the new system design enables a larger range. However, the resolution is degraded by a frequency chirp that appears in the temporal interference signal when a large tilting range is scanned. It is shown through a numerical simulation that the chirp depends on the curvature of the illumination wavefront and also on the position of the pivot axis of the illumination beam. Data processing methods are proposed to overcome these limitations and their effects are illustrated with experimental measurements of opaque surfaces and a weakly scattering phantom with internal features. Displacement measurements involving a controlled rigid body rotation and tilt of a weakly scattering phantom were completed to validate the expected deformations. Both in-plane and out-of-plane components were measured

Methods and Systems for Realizing High Resolution Three-Dimensional Optical Imaging

Methods and systems for realizing high resolution three-dimensional (3-D) optical imaging using diffraction limited low\u27 resolution optical signals. Using axial shift-based signal processing via computer based computation algorithm, three sets of high resolution optical data are detennined along the axial (or light beam propagation) direction using low resolution axial data. The three sets of low resolution data are generated by illuminating the 3~D object under observation along its three independent and orthogonal look directions (i.e., x. Y. and z) or by physically rotating the object by 90 degrees and also flipping the object by 90 degrees. The three sets of high resolution axial data is combined using a unique mathematical function to interpolate a 3-D image of the test object that is of much higher resolution than the diffiaction limited direct measurement 3-D resolution. Confocal microscopy or optical coherence tomography (OCT) are example methods to obtain the axial scan data sets

Roadmap on digital holography [Invited]

This Roadmap article on digital holography provides an overview of a vast array of research activities in the field of digital holography. The paper consists of a series of 25 sections from the prominent experts in digital holography presenting various aspects of the field on sensing, 3D imaging and displays, virtual and augmented reality, microscopy, cell identification, tomography, label-free live cell imaging, and other applications. Each section represents the vision of its author to describe the significant progress, potential impact, important developments, and challenging issues in the field of digital holography

Holographic Digital Fourier Microscopy for Selective Imaging of Biological Tissue

This paper presents an application of digital Fourier holography for selective imaging of scatterers with different sizes in turbid media such as biological tissues. A combination of Fourier holography and high-resolution digital recording, digital Fourier microscopy (DFM) permits crucial flexibility in applying filtering to highlight scatterers of interest in the tissue. The high-resolution digital hologram is a result of the collation of Fourier holographic frames to form a large-size composite hologram. It is expected that DFM has an improved signal-to-noise ratio as compared to conventional direct digital imaging, e.g. phase microscopy, as applied to imaging of small-size objects. The demonstration of the Fourier filtering capacity of DFM using a biological phantom represents the main focus of this paper.Comment: 24 pages, 5 figure